Effect of Leptin Receptor Q223R Polymorphism on Clostridioides difficile Infection-Induced Macrophage Migration Inhibitory Factor Production.

Proinflammatory cytokine levels and host genetic makeup are key determinants of Clostridioides difficile infection (CDI) outcomes. We previously reported that blocking the inflammatory cytokine macrophage migration inhibitory factor (MIF) ameliorates CDI. Here, we determined kinetics of MIF production and its association with a common genetic variant in leptin receptor (LEPR) using blood from patients with CDI. We found highest plasma MIF early after C difficile exposure and in individuals who express mutant/derived LEPR. Our data suggest that early-phase CDI provides a possible window of opportunity in which MIF targeting, potentially in combination with LEPR genotype, could have therapeutic utility.

Development of bioactive and antimicrobial nano-topography over selective laser melted Ti6Al4V implant and its in-vitro corrosion behavior.

Additive manufacturing (AM) or 3D printing of bone defect models is gaining much attention in the biomedical field as it could significantly facilitate the development of customized implants with a high degree of dimensional accuracy. Due to their satisfactory biocompatibility and minimal stress shielding effect, Ti6Al4V (Ti64) alloys are increasingly preferred in the development of such implants. However, their poor osseointegration abilities and lack of antibacterial properties often cause implant loosening and microbial infections, leading to implant failure. To address these drawbacks, we propose in this work a simple surface modification approach of customized Ti64 alloys (3D printed Ti6Al4V) that enables the formation of porous calcium titanate (CT) over their surface as well as the incorporation of silver nanoparticles (AgNPs) into the thus formed porous network. The successful CT formation with the incorporation of AgNPs throughout the 3D printed Ti64 surface and their influence in changing the morphological and mechanical behaviour were studied by Raman spectroscopy, SEM, AFM, Contact angle measurement, XPS, HR-TEM and nano-indentation. Antibacterial studies using Staphylococcus aureus and Escherichia coli, and in-vitro cell studies using MG-63 cell lines showed that surface modified samples resulting from the proposed method exhibit satisfactory antimicrobial property and are highly biocompatible. The obtained surface modified samples also showed a significant improvement in corrosion resistance as compared to unmodified 3D printed Ti64 alloys. The improvement in corrosion resistance was revealed by electrochemical impedance Spectroscopy (EIS). Obtained results emphasis that thus surface modified 3D printed Ti64 alloys are promising candidates for hard tissue implant applications.

Cerium Ion-Incorporated Titanium Metal Implants of Enhanced Bioactivity for Biomedical Applications.

The enhancement in the performance of metallic bone implants based on commercially pure titanium (CP-Ti) by incorporation of cerium (Ce) ions onto the surface was evaluated. The incorporation of Ce ions onto the CP-Ti surface was carried out by a simple two-step chemical treatment method, where an initial NaOH treatment and then a subsequent treatment with different molar concentrations of ceric nitrate solution followed by heat treatment at 600 °C were carried out. The modified surfaces were observed using field emission scanning electron microscopy (FE-SEM), scanning electron microscopy-energy dispersive X-ray analysis (SEM-EDX), X-ray photoelectron spectroscopy (XPS), the laser Raman spectroscopic technique, high-resolution transmission electron microscopy (HR-TEM), and atomic force microscopy (AFM). The formation of a nanonetwork structure by the initial NaOH treatment and the replacement of Na ions with Ce ions along with different phases of TiO2 was evident from the surface characterization results. The transition of rutile TiO2 to anatase TiO2 in the modified surface is evident from the Raman spectra with respect to the treatment of higher to lower concentrations of ceric nitrate solution. The presence of two different oxidation states of Ce (Ce3+ and Ce4+) and improvement in the surface wettability were also distinct in the modified samples. Thus, the incorporated Ce ions over the nanostructured titania network showed low cytotoxicity, good cell adhesion, and enhanced extracellular mineralization on MG-63 cells with better protein adsorption in BSA medium. Taken together, the thus-improved nanostructured surface morphology with the anatase TiO2 phase and distinct extracellular mineralization in the Ce-incorporated Ti metal with good biocompatibility make it a promising candidate for bone implant applications.

Hepatitis B virus genotypes and drug resistance mutations circulating in blood donors in Beira, Mozambique.

Hepatitis B virus (HBV) infects nearly 300 million people and is the leading cause of hepatitis and hepatocellular carcinoma worldwide. Despite the high burden of HBV in sub-Saharan Africa, countries such as Mozambique have limited data available on circulating HBV genotypes and the presence of drug resistance mutations. Blood donors from Beira, Mozambique were tested for HBV surface antigen (HBsAg) and HBV DNA at the Instituto Nacional de Saúde in Maputo, Mozambique. Regardless of HBsAg status, donors with detectable HBV DNA were evaluated for HBV genotype. PCR was performed with primers amplifying a 2.1-2.2 kilobase fragment of the HBV genome. PCR products were submitted for next generation sequencing (NGS), and consensus sequences were evaluated for HBV genotype, recombination, and the presence or absence of drug resistance mutations. Of the 1281 blood donors tested, 74 had quantifiable HBV DNA. The polymerase gene could be amplified from 45 of 58 (77.6%) individuals with chronic HBV infection and 12 of 16 (75%) with occult HBV infection. Among these 57, 51 (89.5%) sequences belonged to HBV genotype A1, while 6 (10.5%) were HBV genotype E. All genotype E sequences were E/A recombinants, and clustered separately from other genotype E references. Genotype A samples had a median viral load of 637 IU/mL, while genotype E samples had a median viral load of 476,084 IU/mL. No drug resistance mutations were observed in the consensus sequences. The current study demonstrates the genotypic diversity of HBV in blood donors in Mozambique, but the absence of dominant (consensus) drug resistance mutations. Studies in other at-risk populations are essential for understanding the epidemiology, risk of liver disease, and likelihood of treatment resistance in resource-limited settings.

Genome Sequence of VanLee, a Singleton Actinobacteriophage That Infects Multiple Gordonia Strains.

VanLee is a singleton phage that was isolated from soil in Florida using Gordonia rubripertincta NRRL B-16540 as the host. The genome is 84,560 bp and has a GC content of 67.8%. VanLee has 164 predicted protein-coding genes and one tRNA. VanLee can infect Gordonia terrae with the same efficiency as G. rubripertincta.

Rotavirus gastroenteritis in Indian children < 5 years hospitalized for diarrhoea, 2012 to 2016.

BACKGROUND: In 2016, the Government of India introduced the oral rotavirus vaccine (ROTAVAC, Bharat Biotech, India) in 4 states of India as part of the Universal Immunization Programme, and expanded to 5 more states in 2017. We report four years of data on rotavirus gastroenteritis in hospitalized children < 5 years of age prior to vaccine introduction. METHODS: Children from 7 sites in southern and northern India hospitalized for diarrhoea were recruited between July 2012 and June 2016. Stool samples were screened for rotavirus using enzyme immunoassay (EIA). The EIA positive samples were genotyped by reverse-transcription polymerase chain reaction. RESULTS: Of the 5834 samples from the 7 sites, 2069 (35.5%) were positive for rotavirus by EIA. Genotyping was performed for 2010 (97.1%) samples. G1P[8](56.3%), G2P[4](9.1%), G9P[4](7.6%), G9P[8](4.2%), and G12P[6](3.7%) were the common genotypes in southern India and G1P[8](36%), G9P[4](11.4%), G2P[4](11.2%), G12P[6](8.4%), and G3P[8](5.9%) in northern India. CONCLUSIONS: The study highlights the high prevalence of rotavirus gastroenteritis in India and the diversity of rotavirus genotypes across different geographical regions. Pre- vaccine surveillance data is necessary to evaluate the potential change in admission rates for gastroenteritis and circulating rotavirus genotypes after vaccine introduction, thus assessing impact.

Direct Medical Costs in Children with Rotavirus and Non-rotavirus Diarrhea Admitted to a Pediatric Intensive Care Unit and High Dependency Unit in Delhi.

OBJECTIVE: To estimate direct medical costs of diarrheal hospitalization of children <5 years admitted in pediatric intensive care unit (PICU) or high dependency unit (HDU). METHODS: Analysis of medical records and hospital bills of 84 children during two time frames, 2005-08 and 2012-14. RESULTS: Direct medical costs in PICU increased from INR 17,941 to INR 50,663 per child for rotavirus diarrhea and INR 11,614 to INR 27,106 for non-rotavirus diarrhea, and in HDU from approximately INR 5,800 to INR 10,500 per child for all-cause diarrhea between the two time frames. CONCLUSIONS: Costs of PICU and HDU care are high and should be included in cost-effectiveness analysis of vaccination.

Multi-center surveillance of rotavirus diarrhea in hospitalized children <5 years of age in India, 2009-2012.

Diarrheal disease due to Group A rotaviruses continues to be an important cause of morbidity in the developing world and India contributes significantly to the disease burden. Surveillance carried out between July 2009 and June 2012 at two medical centers in south India and one center in north India estimated 39% of all diarrheal admissions to be due to rotavirus. The most prevalent genotype isolated was G1P[8](33%) followed by G2P[4](17%). G9P[4] has also emerged as a significant cause of rotavirus diarrhea. No seasonal variation was noticed from the centers in south India, whereas we observed increased rotavirus diarrhea in the center in north India during March and April.

Severity of rotavirus gastroenteritis in an Indian population: report from a 3 year surveillance study.

This study investigated the severity of rotavirus gastroenteritis (RVGE) in hospitalized children less than 60 months of age and compared severity in the first five months of life to severity in children 6 to 23 months of age. Results from a 3 year surveillance study show an early peak of rotavirus disease, with 117 (31%) RVGE hospitalizations in children <6 months old. Higher incidence of severe dehydration, acidemia and acidosis at admission and prolonged hospitalization >7 days were seen in infants 0-5 months of age. The findings support the need for consideration of timely immunization or an accelerated immunization schedule with a birth dose to protect this vulnerable age.

Limb girdle muscular dystrophy due to LAMA2 mutations: diagnostic difficulties due to associated peripheral neuropathy.

We report an eleven year old girl with early motor difficulties initially diagnosed with a peripheral neuropathy in another hospital based on abnormal electrophysiological findings. Our clinical assessment did not highlight obvious clinical features supporting a peripheral neuropathy but evidence of mild proximal weakness. Electrophysiological studies performed at our hospital revealed evidence of a sensorimotor demyelinating polyneuropathy with possible axonal involvement. Brain magnetic resonance imaging (MRI) revealed subtle white matter signal abnormalities, interpreted as nonspecific. Given the patient's proximal weakness and a mildly elevated serum creatine kinase, we performed a muscle biopsy. The muscle had mildly dystrophic features and subtly depleted laminin α2 expression. There was diffusely upregulated laminin α5 expression, and depletion of laminin α2 in intramuscular motor nerves, which made us suspect a partial laminin α2 (merosin) deficiency. Muscle MRI showed predominant posterior and medial compartments involvement. The patient was found to have autosomal recessively inherited double heterozygous LAMA2 mutations. This case illustrates the mild end of the partial merosin deficiency phenotypic spectrum, and highlights how careful assessment of laminin α2 expression in intramuscular motor nerves can be a helpful diagnostic clue in partial merosin deficiency.

Treatable childhood neuronopathy caused by mutations in riboflavin transporter RFVT2.

Childhood onset motor neuron diseases or neuronopathies are a clinically heterogeneous group of disorders. A particularly severe subgroup first described in 1894, and subsequently called Brown-Vialetto-Van Laere syndrome, is characterized by progressive pontobulbar palsy, sensorineural hearing loss and respiratory insufficiency. There has been no treatment for this progressive neurodegenerative disorder, which leads to respiratory failure and usually death during childhood. We recently reported the identification of SLC52A2, encoding riboflavin transporter RFVT2, as a new causative gene for Brown-Vialetto-Van Laere syndrome. We used both exome and Sanger sequencing to identify SLC52A2 mutations in patients presenting with cranial neuropathies and sensorimotor neuropathy with or without respiratory insufficiency. We undertook clinical, neurophysiological and biochemical characterization of patients with mutations in SLC52A2, functionally analysed the most prevalent mutations and initiated a regimen of high-dose oral riboflavin. We identified 18 patients from 13 families with compound heterozygous or homozygous mutations in SLC52A2. Affected individuals share a core phenotype of rapidly progressive axonal sensorimotor neuropathy (manifesting with sensory ataxia, severe weakness of the upper limbs and axial muscles with distinctly preserved strength of the lower limbs), hearing loss, optic atrophy and respiratory insufficiency. We demonstrate that SLC52A2 mutations cause reduced riboflavin uptake and reduced riboflavin transporter protein expression, and we report the response to high-dose oral riboflavin therapy in patients with SLC52A2 mutations, including significant and sustained clinical and biochemical improvements in two patients and preliminary clinical response data in 13 patients with associated biochemical improvements in 10 patients. The clinical and biochemical responses of this SLC52A2-specific cohort suggest that riboflavin supplementation can ameliorate the progression of this neurodegenerative condition, particularly when initiated soon after the onset of symptoms.

Diversity of circulating rotavirus strains in children hospitalized with diarrhea in India, 2005-2009.

BACKGROUND: India accounts for 22% of the 453,000 global rotavirus deaths among children <5 years annually. The Indian Rotavirus Strain Surveillance Network provides clinicians and public health partners with valuable rotavirus disease surveillance data. Our analysis offers policy-makers an update on rotavirus disease burden with emphasis on regional shifts in rotavirus strain epidemiology in India. METHODS: Children <5 years requiring hospitalization for acute gastroenteritis were selected from 10 representative hospitals in 7 cities throughout India between November 2005 through June 2009. We used a modified World Health Organization protocol for rotavirus surveillance; stool specimens were collected and tested for rotavirus using enzyme immunoassay and reverse-transcription polymerase chain reaction. RESULTS: A total of 7285 stool specimens collected were tested for rotavirus, among which 2899 (40%) were positive for rotavirus. Among the 2899 rotavirus detections, a G-type could not be determined for 662 (23%) and more than one G type was detected in 240 (8%). Of 1997 (69%) patients with only one G-type, the common types were G1 (25%), G2 (21%), G9 (13%), and G12 (10%). The proportion of rotavirus infections attributed to G12 infections rose from 8% to 39% in the Northern region and from 8% to 24% in the Western region. CONCLUSIONS: This study highlights the large, ongoing burden of rotavirus disease in India, as well as interesting regional shifts in rotavirus strain epidemiology, including an increasing detection of G12 rotavirus strains in some regions. While broad heterotypic protection from rotavirus vaccination is expected based on pre- and post-licensure data from other settings, effectiveness assessments and rotavirus strain monitoring after vaccine introduction will be important.

Multisite study of cryptosporidiosis in children with diarrhea in India.

Cryptosporidium spp., a common cause of diarrhea in children, were investigated in the first multisite study in India. Diarrheal stools from hospitalized children aged <5 years from Delhi, Trichy, and Vellore were analyzed by microscopy, PCR-restriction fragment length polymorphism (RFLP), and/or sequencing at the small-subunit (SSU) rRNA and Cpgp40/15 loci for species determination and subgenotyping, respectively. Seventy of 2,579 (2.7%) children, 75% of whom were <2 years old, had cryptosporidial diarrhea as determined by microscopy. Genotyping and subgenotyping showed that Cryptosporidium hominis was the most commonly identified species (59/67 children), and subgenotypes Ie, Ia, Ib, and Id were common in all centers. A novel C. parvum subgenotype, IIn, was identified in Vellore. Meteorological analysis revealed a higher rate of cryptosporidial positivity during hotter and drier weather in Delhi.